GLOBAL ANALYSIS OF ANTIBODY REPERTOIRES .1. AN IMMUNOBLOT METHOD FOR THE QUANTITATIVE SCREENING OF A LARGE NUMBER OF REACTIVITIES

This paper describes a procedure for analysing multiple antibody react ivities that explores a commercially available immunoblot system, and is based on a double staining of nitrocellulose membranes, revealing b oth antibody reactivities and the migration position of the blotted pr oteins in the membrane. Quantification of both stainings by densitomet ry allowed the accurate superposition of the immunoreactivity and tota l protein profiles of each lane. Moreover, the protein stainings of th e different lanes could be adjusted with a simple-scale transformation algorithm, correcting for possible distortions during electrophoretic migration, and allowing for the precise comparison of the immunoreact ivity profiles in different lanes. The procedure is discriminatory eno ugh to identify unique reactivity patterns in random pools of 10(4) ac tivated B cells, and to define strain-specific natural antibody repert oires.The utility of this immunoblot method as an assay for simultaneo usly scoring multiple reactivities to hundreds of antigens in complex mixtures of antibodies, and thus defining antibody repertoires in a gl obal manner, is discussed.